LOCUS       sequence-295382-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5304)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133462
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             545..565
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             599..676
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             704..724
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1173..1294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1184..1203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1238..1257
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1409..1455
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1485..1621
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1666..1737
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1963..2551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2043..2062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3297..3365,3366..4157)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3515..3534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4170..4306
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4458..4529
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4534..4580)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4598..4646
                     /note="synthetic polyadenylation signal"
     enhancer        4727..5030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5032..5234
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5185..5205
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5303..5304,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggag tctctcgttc ccggctttaa tgaaaaaact catgtccagt tatccttgcc
       61 agtgcttcaa gtaagggatg tgctggtgcg gggtttcgga gacagtgtag aggaagtcct
      121 gtcagaggct agacagcacc tgaaggatgg gacctgtggc ttagtggaag ttgagaaggg
      181 agtcctcccg cagctggaac aaccttatgt gtttataaaa cgcagcgacg cgcgaacagc
      241 acctcacggg catgtaatgg tggagcttgt cgccgagttg gagggcatcc agtacggaag
      301 atcgggtgaa acgctgggag ttctagtgcc ccacgttggc gaaattccgg tggcctaccg
      361 taaggttttg ctcaggaaaa acggaaataa gggggctggt ggccatagct atggggcaga
      421 cctgaagtct ttcgatctgg gcgacgagct gggcaccgac ccatatgagg atttccagga
      481 aaactggaac acaaaacaca gctccggggt aactcgcgag ctaatgcgag aattgaatgg
      541 aggggaaaac ctttactttc agtcgggacg gcgaatcccg gggctgataa atccctggaa
      601 aaggagatgg aagaagaact ttattgcagt gtctgcggct aaccggttca aaaagatcag
      661 ttcaagcggc gccctagatt acgacattcc tacgaccgcc tccgagaatc tctatttcca
      721 gggtgaattt ggcttggcac aacatgacga ggccgtcgat aacaagttta ataaagaaca
      781 gcagaacgct ttctatgaga tccttcactt accaaatctg aacgaagagc aacgcaacgc
      841 tttcattcag agcctgaaag acgatccctc gcagtctgca aatctcctgg ccgaagccaa
      901 gaagctgaat gacgcacagg ctccaaaagt tgataacaag tttaacaagg agcagcagaa
      961 tgctttctac gagatattgc accttcctaa tctgaatgag gaacaaagaa atgcatttat
     1021 ccagtcccta aaagacgatc catcacagag tgccaacctg cttgcggaag ccaaaaagct
     1081 caatgatgcc caggccccga aagtagacgc gaaccatcaa taagctagct tgactgactg
     1141 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     1201 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1261 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1321 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1381 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     1441 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     1501 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     1561 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     1621 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     1681 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     1741 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     1801 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     1861 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     1921 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1981 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2041 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2101 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2161 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2221 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2281 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2341 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2401 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2461 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2521 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2641 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2701 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2761 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     2821 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     2881 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     2941 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     3001 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     3061 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     3121 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     3181 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     3241 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     3301 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3361 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     3421 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3481 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3541 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3601 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3661 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     3721 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3781 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3841 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     3901 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3961 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4021 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4081 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4141 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     4201 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     4261 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     4321 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     4381 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     4441 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     4501 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     4561 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     4621 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     4681 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4741 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4801 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4861 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4921 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4981 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     5041 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     5101 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5161 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5221 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     5281 ctcgacacac ccgccagcgg ccgc
//
